| Autor: |
Shields TP; Department of Chemistry and Biochemistry, University of Colorado at Boulder 80309-0215, USA., Mollova E, Ste Marie L, Hansen MR, Pardi A |
| Jazyk: |
angličtina |
| Zdroj: |
RNA (New York, N.Y.) [RNA] 1999 Sep; Vol. 5 (9), pp. 1259-67. |
| DOI: |
10.1017/s1355838299990945 |
| Abstrakt: |
An improved method is presented for the preparation of milligram quantities of homogenous-length RNAs suitable for nuclear magnetic resonance or X-ray crystallographic structural studies. Heterogeneous-length RNA transcripts are processed with a hammerhead ribozyme to yield homogenous-length products that are then readily purified by anion exchange high-performance liquid chromatography. This procedure eliminates the need for denaturing polyacrylamide gel electrophoresis, which is the most laborious step in the standard procedure for large-scale production of RNA by in vitro transcription. The hammerhead processing of the heterogeneous-length RNA transcripts also substantially improves the overall yield and purity of the desired RNA product. |
| Databáze: |
MEDLINE |
| Externí odkaz: |
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